Whole-Process of Antibody Drug Development - Facilitate IND Declaration of Biological Drugs

2023-03-03

Page View：

With the rapid development of biopharmaceutical research, various types of biological drugs such as monoclonal antibody, dual antibody, ADC, gene and cell therapy have become an important part of modern biomedicine. Among them, antibody drugs have always occupied the largest share of the global biopharmaceutical market.

Antibody drugs have the advantages of high specificity, rich diversity, uniform properties, long half-life, and can be prepared for specific targets. Antibody drugs have attracted much attention in the treatment of various diseases, especially in the field of therapy.

The R&D of antibody drugs has gone through different development stages such as murine antibodies, chimeric antibodies, humanized antibodies and fully human antibodies. With the increasing variety and wide application of antibody-related drugs, the related immunogenicity issues are gradually exposed to the public view.

❖ Antibody R&D Service

Medicilon provides antibody drugs discovery, CMC research (API + formulation), pharmacodynamics research, PK study, safety evaluation and other services. As of the end of June 2023, Medicilon has successfully assisted in the clinical approval of 31 antibody drugs (8 approved by FDA and NMPA, 1 approved by NMPA and TGA, 1 approved by FDA, NMPA, and TGA) and has multiple antibody projects under development.

01 Anti-Drug antibody(ADA)

Anti-drug antibody (ADA) is the major way to evaluate the immunogenicity of antibody drugs.

There are two mechanisms for the generation of ADA:

One is to use antibody drugs as exogenous protein (such as the first generation of mouse antibodies, chimeric antibodies), similar to immunization with vaccines, to produce a stronger immune response;

The second is that the body's tolerance mechanism of self-proteins (such as fully human antibodies) is destroyed, which is similar to the mechanism of autoantibody production in autoimmune diseases.

Therefore, the formation of ADA can be divided into two major factors: patients and drugs.

Factors related to the formation of ADA.webp

Factors related to the formation of ADA^[1]

Patient Related Factors:

1. The patient's own conditions: Such as low serum albumin concentration, high BMI, genetics, etc

2. Combination medication: If certain immunosuppressants are used in combination, ADA will be reduced.

3. Type of Disease: Under the same medication conditions, patients with rheumatoid arthritis are more likely to develop ADA than patients with spondyloarthritis, which is related to the mechanism of the disease and the impact of the disease on the immune system.

Drug Related Factors

1. Antibody origin: Humanized transformation does not completely eliminate ADA, even if the CDR region of the humanized antibody still has strong immunogenicity, resulting in anti-idiotypic antibodies.

2. Post-translational modification: Studies have shown that removing N-linked glycosylation of the Fc segment can reduce immunogenicity.

3. Preparation process: Formulation, container system, storage conditions may affect immunogenicity through interaction of ingredients with therapeutic protein and container closure system. Process residual impurities such as host DNA and host proteins can also increase ADA production.

4. Dosing regimen: Multiple, high-dose injections are more prone to ADA.

Possible clinical consequences of anti-drug antibodies include decreased drug efficacy, local reactions, and allergic reactions. Not all induced antibodies are present in serum; they may also be present in various organs. Therefore, ADA needs to be tested and evaluated in the antibody development stage to ensure the safety and efficacy of the drug.

02 Introduction of Antibody Development Technology

In the research and development process of developing innovative antibody drugs or antibody-related drugs for a specific target, screening suitable candidate antibodies is a very important procedure. The most common methods for antibody discovery nowadays are hybridoma technology, phage display technology, single B cell antibody screening technology, etc.

Hybridoma Technology

Hybridoma technology, also known as monoclonal antibody technology, is developed on the basis of somatic cell fusion technology. It refers to the fusion of myeloma cells and spleen cells of immunized animals to form highly specific monoclonal antibodies that can secrete immune antigens. Fusion cells not only have the characteristics of unlimited reproduction of tumor cells, but also have the ability of lymphocytes to secrete specific antibodies or cytokines, so that two nuclei from different sources can perform biological functions in the same cell. At the same time, it overcomes the shortcoming of primary lymphocyte proliferation in vitro.

Hybridoma Technology.webp

Hybridoma Technology

Generally speaking, hybridoma technology has the following four steps:

• Immunization: Optimal immunization strategy based on antigen selection by assay design

• Cell fusion: common methods are electrofusion and polyethylene glycol fusion

• Clonal screening: ELISA, IHC, FACS, IFA, etc.

• Antibody production: (in vivo culture) mouse ascites, (in vitro culture) cell supernatant

Phage Display Technology

Phage display technology integrates the gene sequence of exogenous proteins or polypeptides into the phage coat protein gene. In this way, exogenous proteins or polypeptides can be fused into the coat protein for expression, and the exogenous proteins or polypeptides are displayed on the surface of the phage with the biotechnology.

Phage Display Technology.webp

Phage Display Technology ^[2]

Phage display systems are commonly used in the following applications:

• Construction of phage display antibody library (human, rabbit, mouse, alpaca antibody library)

• Construction of phage display peptide library; epitope screening (high-throughput 96*n monoclonal ELISA screening)

• Mature antibody affinity, short preparation cycle, suitable for large-scale industrial production of antibodies

Single B Cell Antibody Screening Technology

Single B cell antibody screening technology is a rapid monoclonal antibody development technology. Only one functional heavy chain variable region gene sequence and one functional light chain variable region gene sequence are expressed in a single B cell gene; therefore, each B cell produces only one specific antibody. The single B cell screening process is to isolate antigen-specific B cells from spleen tissue or peripheral blood of immunized animals. After the isolated single B cell was lysed, the cDNA was obtained by reverse transcription, and the heavy chain and light chain variable region gene sequences of the secreted antibody were amplified by PCR. The monoclonal antibody with potent activity is then expressed in mammalian cells.

03 Medicilon Antibody Development Platform

The Medicilon antibody development platform has been deeply involved in the field of antibody drug research and development and their technical services include the ADA antibody development technical services and druggable antibody development technical services, ADA-free polyclonal antibody development, ADA mouse hybridoma monoclonal antibody development, phage display rabbit monoclonal antibody development services, mouse monoclonal antibody development technical services, mouse/rabbit single B cell antibody development technical services and other projects.

Continuous innovation and breakthrough and self-improvement, Medicilon has summed up years of experience in antibody drug research and development technology, with advanced equipment and condensed into an eye-catching highlight on the antibody development platform:

• Antibody purification: The purity of antigen can be as high as 95% with advanced purification equipment

• Animal immunity: SPF grade experimental animal room with professional and compliant experimental management system

• High-throughput screening:Automatic sampler, robotic arm combined with automatic plate washer to achieve a large number of screening up to 200 plates at a time, which greatly saves time and labor and improves efficiency

• Antibody detection: Advanced ELISA and flow cytometry techniques are used for antibody binding experiments to ensure the delivery standards of antibodies

Medicilon Antibody Development Platform.webp

Medicilon can also provide full-process services for antibody drugs, including CMC research services for antibody drugs, comprehensive preclinical research services in compliance with GLP regulations to help clients obtaining approval with compliant, efficient, and high-quality services. Since the establishment of the platform, it has served many well-known pharmaceutical companies and scientific research institutions. Medicilon has successfully helped the antibody projects of pharmaceutical companies such as Jemincare, Bio-Thera, and CSPC Pharmaceutical Group to be approved for clinical trials. Medicilon reached a long term collaboration with those pharmaceutical giants and won the continuous trust and praise from the clients.

04 Conclusion

After decades of research and development, antibody drugs have made great progress in the field of tumor and autoimmune disease treatment, and have broad development prospects in the field of medicine.

As far as the monoclonal antibody drug market is concerned, it is expected that the global antibody drug will exceed USD 230 billion in 2023, accounting for more than 50% of the global biological products market. The antibody drug approved by the FDA every year accounts for about one-fifth of new drugs approved. All kinds of data show that antibody drugs have become an important part of modern biomedicine.

References:

[1] Anna Vaisman-Mentesh et al，The Molecular Mechanisms That Underlie the Immune Biology of Anti-drug Antibody Formation Following Treatment With Monoclonal Antibodies，Front. Immunol. 11:1951.

[2] Selena Mimmi, et al. Phage Display: An Overview in Context to Drug Discovery. Trends Pharmacol Sci. 2019 Feb;40(2):87-91. doi: 10.1016/j.tips.2018.12.005.

antibody-development

Return

Relevant news

Custom Antibody Development Services

2016-10-19

We provide a complete range of custom monoclonal antibody development services, from antigen design and production to humanized antibodies. We will use an antigen that you supply or we can produce it for you, either as a recombinant protein or a chemically synthesized peptide.

Holiday Promotion: Medicilon Antibody Development Technology Service with Discount Price

2022-12-09

Medicilon Antibody Development Technical Service is here to help you develop new drugs with high cost performance, high efficiency and high quality.